Human Serum Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro and In Vivo

Abstract

Human dental pulp is a promising alternative source of stem cells for cell-based tissue engineering in regenerative medicine, for the easily recruitment with low invasivity for the patient and for the self-renewal and differentiation potential of cells. So far, in vitro culture of mesenchymal stem cells is usually based on supplementing culture and differentiation media with foetal calf serum (FCS). FCS is known to contain a great quantity of growth factors, and thus to promote cell attachment on plastic surface as well as expansion and differentiation. Nevertheless, FCS as an animal origin supplement may represent a potential means for disease transmission besides leading to a xenogenic immune response. Therefore, a significant interest is focused on investigating alternative supplements, in order to obtain a sufficient cell number for clinical application, avoiding the inconvenients of FCS use. In our study we have demonstrated that human serum (HS) is a suitable alternative to FCS, indeed its addition to culture medium induces a high hDPSCs proliferation rate and improves the in vitro osteogenic differentiation. Furthermore, hDPSCs-collagen constructs, pre-differentiated with HS-medium in vitro for 10 days, when implanted in immunocompromised rats, are able to restore critical size parietal bone defects. Therefore these data indicate that HS is a valid substitute for FCS to culture and differentiate in vitro hDPSCs in order to obtain a successful bone regeneration in vivo.

Notch/Wnt cross-signalling regulates stemness of dental pulp stem cells through expression of neural crest and core pluripotency factors

“…In this regard, despite ongoing progress in culture media formulations that do not contain foetal serum for maintenance of DPSCs (Bonnamain et al, 2013;Eubanks et al, 2014;Jung et al, 2016;Xiao and Tsutsui, 2013), currently the addition of FBS or related agents to the culture media permits easily overcoming the issue of initial cell expansion. However, it is becoming increasingly apparent that FBS-containing media also induce differentiation of DPSCs into a default osteo/odontogenic pathway (Pisciotta et al, 2012;Yu et al, 2010) and this may not be the best choice to generate certain cell lineages, particularly neural cells, from DPSCs (Jung et al, 2016). Moreover, upon continual expansion induced by the presence of 10 % FBS, DPSC cultures also tend to generate populations of committed/differentiated cells (Mokry et al, 2010).…”

How has dental pulp stem cells isolation been conducted? A scoping review

“…Serum-free media have been continuously tested for stem cell culture; however, so far, the literature indicates that these media do not adequately support DPSCs proliferation and differentiation. 37,38 The growth of DPSCs with a small percentage of serum enriched with epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) could represent a valid perspective. 39,40 Various human blood derivatives have been proposed as alternatives to animal serum for stem cell culture.…”

“…39,40 Various human blood derivatives have been proposed as alternatives to animal serum for stem cell culture. These include autologous or allogenic human serum, 38 human plasma, 37 and human platelet lysates and their released factors. 4 Pisciolaro and collaborators showed that autologous human serum induces higher CFUs capacity, higher degree of mineralization and less cell damage in DPSCs cells then FBS and allogenic human serum.…”

“…38 Pisciotta and collaborators showed that human serum is an appropriate alternative for FCS, enhancing DPSC proliferation rate and improving in vitro differentiation, and when implanted in immunocompromised rats these cells can restore critical size bone defects. 37 Chen and collaborators showed that when 5% of human platelet lysate was incorporated in normal culture medium the proliferation and mineralization rates were enhanced. 4 Although the literature refers to human platelets as a substitute for FBS, some articles used them in association with FBS, more likely as supplement.…”

Effect of Hypoxia on Stemness and Differentiation of Dental Pulp Derived Stem Cells

“…The petri dishes were then incubated at 37°C overnight. Percentage of cells expressing β-galactosidase was measured in three random fields under inverted phase contrast microscope (Nikon 80i, Japan) [20].…”